Methods and compositions for delivering interleukin-1 receptor antagonist

Patent metadata

Field	Value
Publication number	US-2016017010-A1
Application number	US-201514808828-A
Country	US
Kind code	A1
Filing date	Jul 24, 2015
Priority date	Feb 27, 2008
Publication date	Jan 21, 2016
Grant date	—

Abstract

Methods and compositions generating and using an interleukin-1 receptor antagonist (IL-1ra)-rich solution. Methods for generating and isolating interleukin-1 receptor antagonist include incubating a liquid volume of white blood cells and platelets with polyacrylamide beads to produce interleukin-1 receptor antagonist. The interleukin-1 receptor antagonist is isolated from the polyacrylamide beads to obtain the solution rich in interleukin-1 receptor antagonist. Methods for treating a site of inflammation in a patient include administering to the site of inflammation the solution rich in interleukin-1 receptor antagonist.

First claim

1 . A method for generating a composition rich in interleukin-1 receptor antagonist comprising:
contacting a liquid volume comprising white blood cells with polyacrylamide beads; separating the polyacrylamide beads from the liquid volume to obtain a composition rich in interleukin-1 receptor antagonist.

2 . The method of claim 1 , wherein the liquid volume is whole blood or platelet-rich plasma.

3 . The method of claim 1 , wherein contacting comprises incubating the liquid volume with the polyacrylamide beads for a time from about 30 seconds to about 24 hours.

4 . The method claim 1 , wherein the liquid volume comprises whole blood and contacting comprises mixing the whole blood with an anticoagulant.

5 . (canceled)

6 . The method of claim 1 , further comprising concentrating the liquid volume, including centrifuging whole blood to increase the concentration of white blood cells and platelets relative to whole blood.

7 . The method of claim 6 , wherein centrifuging whole blood comprises:
(i) loading the whole blood into a tube comprising a buoy, configured to reach an equilibrium position upon centrifugation of whole blood in the tube, the equilibrium position being between a first fraction comprising red blood cells and a second fraction comprising white blood cells; and (ii) centrifuging the tube so that the buoy defines an interface between the first fraction and the second fraction.

8 . The method of claim 7 , wherein the tube further comprises:
(a) an isolator operable to define an interface between the second fraction and platelet-poor plasma; and (b) the polyacrylamide beads disposed between the buoy and the isolator.

9 . The method of claim 1 , wherein contacting comprises:
(i) loading whole blood into a separator assembly having a filter operable to trap red blood cells; (ii) centrifuging the separator assembly to separate red blood cells in the filter to form filtered blood comprising a liquid volume of white blood cells; and (iii) incubating the filtered blood with the polyacrylamide beads in a concentrator assembly.

10 . A method for generating a composition rich in interleukin-1 receptor antagonist, comprising:
(1) obtaining whole blood from at least one blood donor; (2) loading the whole blood and an anticoagulant into a tube comprising a buoy disposed in the tube, wherein the buoy is configured to reach an equilibrium position upon centrifugation of the whole blood in the tube, the equililbrium position being between a first fraction comprising red blood cells and a second fraction comprising platelet-rich plasma; (3) centrifuging the tube so that the buoy defines an interface between the first fraction and the second fraction; and (4) collecting the second fraction of platelet-rich plasma; (5) loading the second fraction of platelet-rich plasma into a concentrator assembly including polyacrylamide beads, including incubating the polyacrylamide beads and platelet-rich plasma to form a composition having a concentration of interleukin-1 receptor antagonist greater than that of the whole blood; and (6) rotating the concentrator assembly at centrifugal speeds to separate the interleukin-1 receptor antagonist from the polyacrylamide beads to obtain a composition rich in interleukin-1 receptor antagonist.

11 . The method of claim 10 , where incubating the platelet-rich plasma with the polyacrylamide beads is for a time from about 30 seconds to about 24 hours.

12 .- 15 . (canceled)

16 . A method of treating inflammation in a subject comprising administering to a site of inflammation in the subject a composition rich in interleukin-1 receptor antagonist made according to the process of claim 1 .

17 . The method of claim 16 , wherein the inflammation is associated with osteoarthritis.

18 . The method of claim 16 , further comprising administering fibrinogen, thrombin, and calcium to the site of inflammation.

19 - 24 . (canceled)

25 . The method of claim 16 , wherein the composition rich in interleukin-1 receptor antagonist comprises:
(i) interleukin-1 receptor antagonist at a concentration of at least about 10,000 pg/ml; (ii) soluble Tumor Necrosis Factor Receptor 1 at a concentration greater than the concentration of the sTNF-R1 present in whole blood or plasma; (iii) viable white blood cells; and (iv) at least one growth factor, wherein the total concentration of plasma proteins is at least about 80 mg/ml.

26 . The method of claim 25 , wherein the at least one growth factor comprises platelet-derived growth factor-AB, transforming growth factor β1, insulin-like growth factor-1, basic fibroblast growth factor, or vascular endothelial growth factor.

27 - 38 . (canceled)

39 . The method of claim 1 , further comprising filtering whole blood or platelet-rich plasma to obtain the liquid volume comprising white blood cells.

40 . (canceled)

41 . The method of claim 1 , wherein the liquid volume comprises whole blood, and wherein contacting comprises filtering the whole blood and the polyacrylamide beads to remove red blood cells.

42 . (canceled)

43 . The method of claim 1 , wherein the liquid volume is whole blood or plasma, and wherein the composition comprises interleukin-1 receptor antagonist at a concentration of at least about 10,000 pg/ml and soluble Tumor Necrosis Factor Receptor 1 (sTNF-R1) at a concentration greater than the concentration of the sTNF-R1 present in the whole blood or plasma.

44 . The method of claim 1 , wherein the polyacrylamide beads are dry.

45 . The method of claim 1 , wherein contacting comprises allowing the polyacrylamide beads to absorb a portion of the liquid volume.

Assignees

• Biomet Biologics Llc

Inventors

• Higgins Joel C
• Woodell-May Jennifer E
• Hoeppner Jacy C

Classifications

• A61K47/34

  Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers (A61K47/10 takes precedence) · CPC title

• A61K45/06

  Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca · CPC title

• C07K1/34

  by filtration, ultrafiltration or reverse osmosis · CPC title

• A61K38/1709

  from mammals · CPC title

• C07K14/47 Primary

  from mammals · CPC title